Abstract |
The prion protein (PrP) is synthesized in three topologic forms at the endoplasmic reticulum. (sec)PrP is fully translocated into the endoplasmic reticulum lumen, whereas (Ntm)PrP and ( Ctm)PrP are single-spanning membrane proteins of opposite orientation. Increased generation of ( Ctm)PrP in either transgenic mice or humans is associated with the development of neurodegenerative disease. To study the mechanisms by which PrP can achieve three topologic outcomes, we analyzed the translocation of proteins containing mutations introduced into either the N-terminal signal sequence or potential transmembrane domain (TMD) of PrP. Although mutations in either domain were found to affect PrP topogenesis, they did so in qualitatively different ways. In addition to its traditional role in mediating protein targeting, the signal was found to play a surprising role in determining orientation of the PrP N terminus. By contrast, the TMD was found to influence membrane integration. Analysis of various signal and TMD double mutants demonstrated that the topologic consequence of TMD action was directly dependent on the previous, signal-mediated step. Together, these results reveal that PrP topogenesis is controlled at two discrete steps during its translocation and provide a framework for understanding how these steps act coordinately to determine the final topology achieved by PrP.
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Authors | S J Kim, R Rahbar, R S Hegde |
Journal | The Journal of biological chemistry
(J Biol Chem)
Vol. 276
Issue 28
Pg. 26132-40
(Jul 13 2001)
ISSN: 0021-9258 [Print] United States |
PMID | 11359769
(Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)
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Chemical References |
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Topics |
- Amino Acid Sequence
- Animals
- Evolution, Molecular
- Humans
- Molecular Sequence Data
- Mutation
- Prions
(genetics)
- Sequence Alignment
- Sequence Analysis
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